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    Review: What Three Types of Hemolysis Can Be Observed On A BAP Plate?

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    Jonille Echevarria
    I. This lecture discusses biochemical tests used to identify Gram-positive and Gram-negative bacteria, including blood agar plates, mannitol salt agar, antibiotic disks, and fermentation tests. II. Key characteristics of Enterobacteriaceae and Pseudomonas bacteria are outlined, including their environments and representative species like E. coli, Salmonella, and P. aeruginosa. III. Various biochemical tests are described, such as fermentation broth tubes, SIM tubes, MacConkey agar, and Kligler's iron agar, which are used to determine properties like sugar fermentation, gas production, and enzyme activity.

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    Review: What Three Types of Hemolysis Can Be Observed On A BAP Plate?

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    Jonille Echevarria
    34 views5 pages
    I. This lecture discusses biochemical tests used to identify Gram-positive and Gram-negative bacteria, including blood agar plates, mannitol salt agar, antibiotic disks, and fermentation tests. II. Key characteristics of Enterobacteriaceae and Pseudomonas bacteria are outlined, including their environments and representative species like E. coli, Salmonella, and P. aeruginosa. III. Various biochemical tests are described, such as fermentation broth tubes, SIM tubes, MacConkey agar, and Kligler's iron agar, which are used to determine properties like sugar fermentation, gas production, and enzyme activity.

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    I. This lecture discusses biochemical tests used to identify Gram-positive and Gram-negative bacteria, including blood agar plates, mannitol salt agar, antibiotic disks, and fermentation tests. II. Key characteristics of Enterobacteriaceae and Pseudomonas bacteria are outlined, including their environments and representative species like E. coli, Salmonella, and P. aeruginosa. III. Various biochemical tests are described, such as fermentation broth tubes, SIM tubes, MacConkey agar, and Kligler's iron agar, which are used to determine properties like sugar fermentation, gas production, and enzyme activity.

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    34 views5 pages

    Review: What Three Types of Hemolysis Can Be Observed On A BAP Plate?

    Uploaded by

    Jonille Echevarria
    I. This lecture discusses biochemical tests used to identify Gram-positive and Gram-negative bacteria, including blood agar plates, mannitol salt agar, antibiotic disks, and fermentation tests. II. Key characteristics of Enterobacteriaceae and Pseudomonas bacteria are outlined, including their environments and representative species like E. coli, Salmonella, and P. aeruginosa. III. Various biochemical tests are described, such as fermentation broth tubes, SIM tubes, MacConkey agar, and Kligler's iron agar, which are used to determine properties like sugar fermentation, gas production, and enzyme activity.

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    of 5

    Lecture 9

    I. Biochemical testing for Gram positive organisms continued


    A. Blood Agar Plates (BAP) review

    Review: What three types of hemolysis can be observed on a BAP plate?


    _alpha, beta and gamma_

    B. MSA plate review


    Review: What color would an MSA plate be if the organism streaked on it was
    able to ferment mannitol?
    Yellow
    ________________________________________________________

    C. Taxos testing (antibiotic resistance)


    1. Tacos P optochin sensitivity testing is a disk inoculated with organisms sensitive to the antibiotic
    optochin and those not.  This antibiotic is used to differentiate between α-hemolytic strains.
    i. Nonpathogenic normal flora, S. mitis, shows optochin sensitive
    ii. Pathogenic S. pneumonia is optochin sensitive and will show a large zone of inhibition.
    2. Tacos A (Bacitracin sensitivity testing) is a disk inoculated with organisms sensitive to the
    antibiotic bacitracin and those not. This is an antibiotic used to differentiate between β-
    hemolytic strains.
    i. Streptococcus pyogenes is bacitracin sensitive
    ii. Streptococcus agalactiae is bacitacin resistant

    II. Gram Negative Organisms


    A. ENTEROBACTERIACEAE 
    Key
    characteristics: Environments
    • Straight, Gram-negative rods Inhabit the gut (“entero” means gut).
    • ___________________ (This is the best way
    to differentiate between Enterobacteriaceae and
    Pseudomonas spp).
    • Non-fastidious (simple nutritional requirements)
    • ______________________
    • If motile, flagella are pertitrichous
    Representative species
    Escherichia coli Proteus mirabilis Shigella dysenteriae
    Known for Characteristics Known for Characteristics Known for Characteristics
    • opportunistic
    • Some strains • ferments pathogen • ferments glucose • causes • ferments
    normal gut
    flora, fecal
    indicator,
    can be an
    opportunisti • swarming
    of E. coli can c pathogen_ Pathogen motility shigellosis glucose, and
    Cause causing UTIs • ______________ mannitol
    diseases such
    as meningitis
    ___ • reduces nitrate
    • reduces • utilizes citrate • motile
    • hydrolyzes urea
    nitrate to ammonia • produces H2S
    . • Produces • utilize citrate
    indole • MR+ •MR+
    • MR+
    Salmonella typhi Klebsiella pneumoniae
    Known for Characteristics Known for Characteristics
    • _ typhoid fever,
    gastroenteritis
    ________________ • ferments_sucrose __ • normal flora • ferments
    • causes lactose
    pneumonia,
    bloodstream
    infections, wound
    or surgical site
    infections, and
    • reduces nitrate meningitis.
    • MR+
    in alcoholics and • reduces nitrate
    people with • utilizes citrate
    compromised • VP+
    pulmonary
    function

    B. PSEUDOMONAS
    Key
    characteristics: Environments
    Opportunisti
    • Straight, or slightly curved Gram-negative rods Generally found pathogen
    • Oxidase positive
    • Non-fermentative
    • Strict aerobes
    • Motile via polar flagella
    • Often produce water soluble pigments and colonies appear mucoid
    Representative species
    P.
    fluorescen
    P. aeruginosa s
    Known for Characteristics Known for Characteristics
    •potential
    • the most clinical significance that causes agents for the
    disease in • reduces nitrate biocontrol • growth temperature
    • produces several
    enzymes and
    immunocompromised (e.g. cystic fibrosis toxins below 30 C
    patients, AIDS patients, and burn victims)
    • produces several enzymes and toxins that
    account for its virulence. The most important
    toxin blocks protein synthesis in eukaryotic
    cells.
    Even the water-soluble pigment pyocyanin
    causes tissue damage to a host
    III. Biochemical testing for Gram negative
    organisms A. Fermentation Broth tubes
    1. These tubes contain a simple sugars in addition to a lactose and a mannitol A positive reaction will
    turn the broth yellow. The Durham tube will measure detect production of gas by the bacteria (gas
    produced by an enzyme called  formic hydrogen lyase .

    2. Glucose tubes
    i. In today’s lab we will be using a glucose broth tube. If the liquid in the glucose tube is acidic, then
    the pH indicator turns yellow. If the liquid in the glucose tube is basic, then the pH indicator will
    remain
    ii. A bubble in the Durham tube indicates that gas has been produced. Many members of the
    family Enterobacteriaceae can metabolize __ which can be converted to formic acid and formic
    acid to H2 (g) and CO2 (g) by an enzyme called formic hydrogen lyase.
    B. SIM Tube (Sulfur, Indole, Motility)
    1. Sulfur: H2s production will turn the agar into black color forms.
    i. H2S is produced either through the catabolism by an enzyme called cysteine desulfurase or by the
    enzyme cysteine desulfurase during anaerobic respiration.
    2. Indole: Adding Kovac’s reagent, and form rosindole dye which is red in color (indole +) determine
    indole production.
    i. Indole is produced from the breakdown of the amino acid cysteine.
    3. Motility: Motility is determined by observing the organism’s ability to move through the
    growth/turbidity away from the line of inoculation.
    C. MacConkey agar
    1. Selective medium
    i. Bile salts inhibit the growth of gram-positive bacteria organisms.
    ii. If the bacteria are able to ferment lactose, the acid produced will cause the pH indicator to dark
    pink/red
    D. Kligler’s Iron Agar (KIA)
    1. A differential medium that contains two sugars: lactose (low concentration) and glucose (higher
    concentration).
    i. Fermenters will typically utilize the manor to test for various fermentation capabilities and
    production of acidic fermentation byproducts will cause the entire tube to turn yellow. However, the
    glucose is in short supply and, after the first few hours of growth, will all be used up.
    ii. After the glucose is gone, bacteria that are capable of fermenting lactose will do so.
    Production of acidic byproducts will continue and the medium will remain yellow.
    iii.Gas produced as a product of fermentation will
    form fissures or completely the agar will be
    lifted off the bottom of the tube.
    iv. Bacteria that are incapable of fermenting lactose
    have to utilize the _____________________ in the
    medium. This produces NH3 (a weak base), which
    causes  the medium to become alkaline. and
    turns the slant red. The butt remains yellow.
    v. Non-fermenters will utilize only the amino acids
    and proteins in the medium. The slant will be red
    and the butt will remain the original red/orange
    color.
    vi. The production of H2S results in a positive for
    glucose fermentation . If the black precipitate
    obscures the medium color, it is
    positive for glucose fermentation.

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